Background staining or non-specific staining is a problem often encountered in immunohistochemistry, immunofluorescence, and in situ staining. Background staining is caused by a number of factors such as the cross-reactivity of antibodies with shared epitopes in the tissue, by the presence of natural antibodies and/or contaminants present in the primary antibody and/or the secondary antibody, by ionic interactions, due to the presence of carbohydrates and endogenous biotin present in the tissue. Background eradication is most important to obtain specific background staining to facilitate qualitative and quantitative evaluation.
Background Buster is a peptide blocker that eradicates all general background staining. Background Buster removes all background staining caused by primary antibodies, staining reagents, chromogens, fixatives, and endogenous biotin present in tissues such as the liver, spleen, and kidney. Background Buster is used in place of normal sera and other blocking solutions to remove background staining in human and animal tissues.
Background Buster is applicable to IHC staining, immunofluorescence staining, and in situ probe staining in both human and animal tissues. It is also applicable to flow cytometry assays.
Background Buster is essential for the staining of animal tissues, it is especially essential when staining tissues and antibodies of identical species, such as mouse antibodies in mouse tissues (mouse on the mouse) and rabbit on the rabbit. A 30-minute incubation with Background Buster is recommended prior to primary antibody application for staining of antibodies and primary tissues of identical species. Use of Background Buster is strongly recommended for staining of indirect species (tissue/antibody from non-identical species) such as Rat-On-Mouse, Mouse-on-Rat, Mouse-on-Rabbit, etc. A 20-minute incubation with Innovex Background Buster is recommended prior to application of the primary antibody for Indirect staining (antibodies and primary tissues of non-identical species).
In immunoperoxidase-IHC staining, red blood cell staining causes another type of background and non-specific staining; This is due to the endogenous peroxidase enzyme present in red blood cells. This type of background requires a pretreatment step with hydrogen peroxide (H2O2) freshly made at 3% in water, this blocking step must precede the blocking step with Innovex Background Buster.